Process for the removal of inert substances from blood-serums, &amp;c.



UNITED STATES PATENT OFFICE.

JULIUS THEODOR ALEXANDER SAMES, OE DUSSELDORF, GERMANY.

PROCESS FOR THE REMOVAL OF INERT SUBSTANCES FROM BLOOD-SERUMS, &c. g

No Drawing.

ments in Processes for the Removal of Inert Substances from Blood-Serums, &c., of which the following is a speclfication;

This invention relates to a process for the removal of inert substances from blood' serums, milk or other liquids derived from animals of various kinds and containing antisubstances or bodies of an enzym nature and the invention is directed mainly to the problem of the direct production of sterile and clear solutions of. anti substances poo-r in albu mi'n. For thisipu'rjpose it"isnne'cessary to remove from'the blood-'h'serunis d rived from annuals as great a portion as; posslble of the albumin which is injurious to the foreign human or animal organism when assimilated, and also any' subsidiary substances.-

The new process consists substantially in treating fresh therapeutic or anti serums, or those which havenot'yet been preserved or have been again freed from preservative agents, with suitable chemicals which are only slightly dissociated such as weakly acid organic precipitants and heating them,

whereby the substances (such as albumin) which do-not contain the ant-itoxin or the anti-substances are precipitated, after which any chemicals still present 1n excess 1n the solution which is now poor in albumin but;

contains, the main portion of the anti-substances and can be easilyconcentrated, are removed by means of suitable compounds in an insoluble form or rendered innocuous,

so that clear and sterile concentrated thera peutic and anti serums are obtained. The

"use of heat simultaneously prevents the forthe anti-substance which is clear and easily maintained in a sterile condition being so obtained. V

In order to obtain precipitates substantially free from antitoxins the serums obtained from horses, or animals of the same family, are preferably made alkaline before being treated. The sterile and clear serum Specification of Letters Patent.

Patented Apr. 1, 1913.

Application filed a rn 9,1912. Serial No. 689,593.

obtained-according to the new process contains the greater portion of the antitoxin,

but only about 50% of, the albumin of the original serum and can be used either direct as a therapeutic serum poor in albumin or, after evaporation in cameo, as a serum containing a very considerably increased amount of immune substances or as a dry serum. I

The treatment according to the new prooess is intended to apply principally to freshly obtained serums, theamount of anti- .sub'stances contained in which has not yet diminished by standing or by preservation. Serums preserved bymeans' of phenol or cresol or their derivatives may howeveralso be treated by this process, but the heating must then be omitted or the" preservative agents mus be removed by repeated extraction with clilorofonn---for example, before applying the process.

An example of the way in which vthe new process is carried out is described in the following example :100 cc. of freshly obtained tetanus serum of a strength of 8 and from horse blood are mixed with 150 cc. of sterile'distil led water and 1 cc. of 10% sodium carbonate solution, to which about 1% of ammonium carbonate has been added, and heated to from 50 to 60 degreescentigrade for 15 minutes in a glass cylinderv fitting into the centrifugal machine. 10 cc. of aluminium acetate solution, of a strength 7.58% of basic aluminium acetate are then added drop by drop to the alkaline liquid with stirring whereby a thick, flocculent and readily deposited precipitate, consisting of aluminium hydroxid and horse albumin, is formed and which is allowed to settle before pouring off the now acid liquid containing the antito-xin, then subsequently centrifuged off and neglected. The ,liquid thus .freed from about half of the horse albumin is made faintly alkaline with the above-men tioned alkalizing agent and then evapo rated down in 'vacuo at a temperature of from 4,0 to 50 degrees. down to cc. the preparation prepared from the tetanus serum of a strength of 8 is shown by chemical, determination to be freed from, roughly, 50% of the albumin.

According to biological tests onwhite mice it was of a strength of 7% and of a clear bright appearance and perfectly sterile.

Owing to the so greatly reduced abumin When evaporated content further evaporation inoacuo for the purpose of increasing the antitoxin content.

which possesses a germicidal action is to bev regarded as an advantage of the process. The use oi" heat and precipitation which carries down any germs with it also eifect sterilization, so that the process can also be used as a sterilizing agent for slightly contaminated therapeutic serums. Old preserved serums, which no longer contain the given number of antito-Xin units, can also be regenerated and intensified again.

Instead of aluminium salts other metallic compounds may also be used for the precipitation of the albumin from horse blood serum. In addition to being used for blood serums obtained from horses or animals of the same family, such as the mule or hinny, the new process may also be used in an an pressly' modified form for blood serums from other kinds of animals such as monkeys, oxen, sheep, goats, pigs, rabbits, guineapigs, &c., which contain reaction products on toxins and infections,,&c., such as lysins, precipitins, agglutinins, complement forming anti-substances, enzyms and similar substances. It is not always necessary in this process to obtain precipitation by metallic hydroxid precipitates produced in a colloidal form, on the contrary, other chemicals may also be 'used for recipitation such as acetic acid or other innocuous organic acids for ox blood serum for example. Finally milk or other liquids de-' rived from animals and containing antisubstances or substances of an enzym nature may alsobe treated by the new process.

When those albumin substances which are distinguished by coarse flocculent precipita tion and thereby indicate that they do not contain the anti-substances have 'been removed by the new process, the albumin which contains the anti-substance can obviously also be separated by a further application of the process. In this case, however, the precipitation is not coarsely fl cculent but finely flocculent.

In the useof the new process in science it is advisable to determine first with small quantities of serum with what quantities of the added chemicals the most advantageous action is obtained for the particular serum under treatment.

Claimsx 1. Process for the removal of inert substances from liquids derived from animal i and similar bodies,

bodies and containing anti-substances, toxins and providing directly sterile and clear'solutions of such anti-substances, which comprises treating the same with a sufficient amount of a weakly acid organic precipitant to precipitate substantially all and only the portion of the albumin of said liquidsnot containing the active principle.

2. Process for the regeneration of, and the removal of inert substances from, liquids derived from animal bodies and containing anti-substances, toxins and similar bodies, and providing directly sterile and clear solutions of such anti-substances, which comprises removing the preservative fromthe liquids; heating said liquids; and treating the same with a sufiicient amount of a suitable organic precipitant to precipitate substantially all and only the portion of the albumin of said liquids not containing the active principle. v

3. Process for the removal of inert substances froin liquids derived from animal bodies and containing anti-substances, toxins and similar bodies, and providing directly sterile and clear solutions of such anti-substances, which comprises adding an alkaline reagent thereto, and treating with a sufficient amount of a salt of a metal forming insoluble hydroxids to precipitate substantially all and only the portion of thel albumin of said liquids not containing the active principles.

4. Process for the removal of inert substances. from liquids derived from animal bodies and containing anti-substances, toxins and similar bodies, and roviding directly sterile and clear solutions .of such anti-substances, which comprises adding-a carbonate of an alkali and treating with a suflicient amount of a salt of a metal forming insoluble hydroxids to precipitate substantially all and only the portion of the albumin of said liquids not containing-the active principles.

5. Process for the removal of inertsubstances from liquids derived from animalbodies" and containing anti-substances, toxinsand similar bodies, and providing directly sterile and clear solutions of such anti-substances, which comprises adding a carbonate of an alkali; and treating with a suflicient amount of an aluminium salt to precipitate substantially all and only the portion of the albumin of said liquids' not containing the active principles.

6. Process for the removal of inert substances from liquids derived from animal bodies and containing anti-substances, toxins and similar bodies, and providing directly sterile and clear solutions of such anti-substances, which comprises adding a carbonate of an alkali; treating with a sufiicient amount of an aluminium salt to pre cipitate substantially all and only the portion of the albumin of said liquids not containing the active principles; and finally adding a carbonate of an alkali and car-- bonate of ammonium for the purpose of s parating insoluble aluminium compounds.

7. Process for the removal of inert suband removing the precipitate formed therestances from tetanus serum, of a strength of by, and finally rendering the solution faintly 8 and obtained from horse. blood, which alkaline;

consists in adding to 100 cc. of such serum In Witness whereof I have hereunto set 5 mixed with 150 cc. of sterile, distilled water, my hand in presence of two Witnesses. 1 cc. of 10% sodium carbonate solution to 7 which about 1% of ammonium carbonate JULIUSTHEODOR ALEXANDER SAMES' has been added, heating the same for fifteen WVitnesses: minutes from 50 C. to 60 (3., then adding HELEN NUF R, 10 thereto 10 cc. of aluminium acetate solution, ALBERT NUFER.

Copies of thi5 patent may be obtained for five cents each, by addressing the Commissioner of Patents,

Washington, D. c. 

